LOCUS       Exported               10759 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Binary vector pCMIRT15, complete sequence.
ACCESSION   HQ540318
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10759)
  AUTHORS   Zhang C, Ng DW, Lu J, Chen ZJ.
  TITLE     Roles of target site location and sequence complementarity in 
            trans-acting siRNA formation in Arabidopsis
  JOURNAL   Plant J. 69 (2), 217-226 (2012)
  PUBMED    21910773
REFERENCE   2  (bases 1 to 10759)
  AUTHORS   Zhang C, Chen ZJ.
  TITLE     Direct Submission
  JOURNAL   Submitted (03-NOV-2010) Section of Molecular Cell and Developmental 
            Biology, The University of Texas at Austin, Austin, TX 78712, USA
REFERENCE   3  (bases 1 to 10759)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..10759
                     /organism="Binary vector pCMIRT15"
                     /mol_type="other DNA"
                     /db_xref="taxon:1033798"
     CDS             complement(34..1059)
                     /codon_start=1
                     /product="hygromycin phosphotransferase"
                     /label=hygromycin phosphotransferase
                     /note="HPT"
                     /protein_id="AEG42343.1"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRSQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     K"
     misc_feature    complement(1060..1444)
                     /note="promoter and 5 UTR of mannose synthase; MAS 
                     promoter"
     promoter        complement(1064..1444)
                     /label=MAS promoter
                     /note="mannopine synthase promoter (Velten et al., 1984)"
     misc_feature    1445..2337
                     /note="35S promoter; CaMV 35S promoter with two copies of 
                     enhancers and tobacco etch virus translation leader 
                     sequence"
     promoter        1852..2196
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     CDS             2338..3528
                     /codon_start=1
                     /product="HTA6-sGFP fusion protein"
                     /label=HTA6-sGFP fusion protein
                     /note="sGFP was modified by the Sheen Lab for enhanced 
                     fluorescence; fusion protein between Arabidopsis core 
                     histone HTA6 and sGFP"
                     /protein_id="AEG42342.1"
                     /translation="MESTGKVKKAFGGRKPPGAPKTKSVSKSMKAGLQFPVGRITRFLK
                     KGRYAQRLGGGAPVYMAAVLEYLAAEVLELAGNAARDNKKSRIIPRHLLLAIRNDEELG
                     KLLSGVTIAHGGVLPNINSVLLPKKSATKPAEEKATKSPVKSPKKAHVGPMVSKGEELF
                     TGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTFTYG
                     VQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELK
                     GIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVNFKIRHNIEDGSVQLADHYQQN
                     TPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEFVTAAGITHGMDELYKGIL"
     CDS             2800..3516
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     3UTR           3529..3909
                     /note="contains octopine synthase terminator, a synthetic 
                     poly linker, and CaMV 35S polyA signal"
     terminator      3925..4177
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     misc_feature    3928..10502
                     /label=from pCAMBIA1302
                     /note="from pCAMBIA1302"
     misc_feature    4199..4223
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     misc_feature    4218..4243
                     /label=right border repeat sequence from C58 T-DNA
                     /note="right border repeat sequence from C58 T-DNA"
     CDS             5523..6152
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             6586..7653
                     /codon_start=1
                     /product="replication protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 RepA
                     /translation="GRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESWQA
                     AADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLSKR
                     DRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKPGR
                     VFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGEAL
                     ISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYRLA
                     RRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPILV
                     MRYRNLIEGEASAGS"
     rep_origin      complement(6877..7877)
                     /direction=LEFT
                     /label=pVS1
                     /note="pVS1"
     rep_origin      7719..7913
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     misc_feature    8257..8397
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(8583..9171)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     rep_origin      complement(8687..8967)
                     /direction=LEFT
                     /label=pBR322
                     /note="pBR322"
     CDS             complement(9258..10052)
                     /codon_start=1
                     /gene="aadA"
                     /product="kanamycin resistance protein"
                     /label=aadA
                     /protein_id="AEG42344.1"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     gene            complement(9258..10052)
                     /gene="aadA"
                     /label=aadA
     misc_feature    10477..10502
                     /label=left border repeat sequence from C58 T-DNA
                     /note="left border repeat sequence from C58 T-DNA"
     misc_feature    10477..10501
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     3UTR           complement(10569..33)
                     /label=contains 35S polyA signal
                     /note="contains 35S polyA signal"
     polyA_signal    10579..10753
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"